Interpretive Summary: Babesia bovis is a tick borne apicomplexan pathogen that remains an important constrain for the development of cattle industries worldwide. The disease can be produced by distinct B. bovis stains with different levels of virulence and other characteristics. This manuscript describes a simple method for the differentiation of such strains occurring in the Americas.

Technical Abstract:
The merozoite surface antigen-2 (msa-2) family of Babesia bovis is a group of variable genes that share conserved 5' and 3' ends and encode for membrane-anchored glycoproteins that have been postulated as vaccine candidates. In this work, we analyzed
the sequences of three of these genes (msa-2a1, a2, and 2b) from two geographically distant strains and detected a certain degree of genotypic diversity that could be exploited to work out new molecular tools for the discrimination of B. bovis field samples.
Here we describe a PCR restriction assay that was developed based on this observation and tested on several B. bovis strains and isolates. The results show a strain-specific band pattern in geographically distant isolates, indicating the presence of differentially located BspMI restriction sites. This approach provides a simple method for the differentiation of American B. bovis strains.